Rapid methodology for antigenic profiling of FMDV field strains and for the control of identity, purity and viral integrity in commercial virus vaccines using monoclonal antibodies

Seki, Cristina; Robiolo, Blanca; Periolo, Osvaldo; Iglesias, Marcela; D'antuono, Alejandra Lorena; Maradei, Eduardo; Barros, Virginia; Torre, J.L. La; Mattion, Nora

doi:10.1016/j.vetmic.2008.07.011

Cited by 27 publications

(39 citation statements)

References 32 publications

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“…This preparation was further centrifuged at 100,000 ϫ g for 16 h at 4°C, and pelleted virus was resuspended in NET buffer, quantified by spectrophotometry at 260 nm, and stored at Ϫ80°C. The integrity of 140S particles was assessed by SDG and a monoclonal antibody (MAb)-based indirect ELISA applied to each gradient fraction, as described previously (22).…”

Section: Methodsmentioning

confidence: 99%

Early Adaptive Immune Responses in the Respiratory Tract of Foot-and-Mouth Disease Virus-Infected Cattle

Pega

Bucafusco

Giacomo

et al. 2013

J Virol

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Foot-and-mouth disease (FMD) is a highly contagious viral disease which affects both domestic and wild biungulate species. This acute disease, caused by the FMD virus (FMDV), usually includes an active replication phase in the respiratory tract for up to 72 h postinfection, followed by hematogenous dissemination and vesicular lesions at oral and foot epithelia. The role of the early local adaptive immunity of the host in the outcome of the infection is not well understood. Here we report the kinetics of appearance of FMDV-specific antibody-secreting cells (ASC) in lymphoid organs along the respiratory tract and the spleen in cattle infected by aerosol exposure. While no responses were observed for up to 3 days postinfection (dpi), all animals developed FMDV-ASC in all the lymphoid organs studied at 4 dpi. Tracheobronchial lymph nodes were the most reactive organs at this time, and IgM was the predominant isotype, followed by IgG1. Numbers of FMDV-ASC were further augmented at 5 and 6 dpi, with an increasing prevalence in upper respiratory organs. Systemic antibody responses were slightly delayed compared with the local reaction. Also, IgM was the dominant isotype in serum at 5 dpi, coinciding with a sharp decrease of viral RNA detection in peripheral blood. These results indicate that following aerogenous administration, cattle develop a rapid and vigorous genuine local antibody response throughout the respiratory tract. Time course and isotype profiles indicate the presence of an efficient T cell-independent antibody response which drives the IgM-mediated virus clearance in cattle infected by FMDV aerosol exposure.

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Section: Methodsmentioning

confidence: 99%

Early Adaptive Immune Responses in the Respiratory Tract of Foot-and-Mouth Disease Virus-Infected Cattle

Pega

Bucafusco

Giacomo

et al. 2013

J Virol

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“…For vaccine matching purpose, reactivity value (Rx) obtained with each mAb for every field isolate and the reference strain (O1/Manisa) was used to calculate r values to normalize the results [16,19], since the O1/Manisa mAb panel was not available. The r values (Table 2) were used to determine the strength of the linear association between the vaccine strain and field isolates.…”

Section: Resultsmentioning

confidence: 99%

“…Control mAbB ) ×100]. The r values between O1/Manisa and each isolate were calculated based on the reactivity (Rx) obtained with each mAb as described by Seki [16]. …”

Section: Methodsmentioning

confidence: 99%

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Characterization of monoclonal antibodies against foot-and-mouth disease virus serotype O and application in identification of antigenic variation in relation to vaccine strain selection

Yang

Goolia

et al. 2014

Virol J

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BackgroundFoot-and-mouth disease (FMD) has severe implications for animal farming which leads to considerable financial losses because of its rapid spread, high morbidity and loss of productivity. For these reasons, the use of vaccine is often favoured to prevent and control FMD. Selection of the proper vaccine is extremely difficult because of the antigenic variation within FMDV serotypes. The aim of the current study was to produce a panel of mAbs and use it for the characterization of new isolates of FMDV serotype O.ResultsA panel of FMDV/O specific mAb was produced. The generated mAbs were then characterized using the peptide array and mAb resistant mutant selection. Seven out of the nine mAbs reacted with five known antigenic sites, thus the other two mAbs against non-neutralizing sites were identified. The mAbs were then evaluated by antigenic ELISA for the detection of forty-six FMDV serotype O isolates representing seven of ten known topotypes. Isolates ECU/4/10 and HKN/2/11 demonstrated the highest antigenic variation compared to the others. Furthermore, the panel of mAbs was used in vaccine matching by antigenic profiling ELISA with O1/Manisa as the reference strain. However, there was no correlation between vaccine matching by antigenic ELISA and the gold standard method, virus neutralisation test (VNT), for the forty-six FMDV/O isolates. Nine isolates had particularly poor correlation with the reference vaccine strain as revealed by the low r1 values in VNT. The amino acid sequences of the outer capsid proteins for these nine isolates were analyzed and compared with the vaccine strain O1/Manisa. The isolate ECU/4/10 displayed three unique amino acid substitutions around the antigenic sites 1, 3 and 4.ConclusionsThe panel of mAbs is useful to monitor the emergence of antigenically different strains and determination of relevant antigenic site differences. However, for vaccine matching VNT remains the preferred method but a combination of VNT, antigenic profiling with a panel of mAbs and genetic sequencing would probably be more ideal for full characterization of any new outbreak isolates as well as for selection of vaccine strains from FMDV antigen banks.

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“…Structural protein characterization was carried out by ELISA using four different monoclonal antibodies (MAbs 1-5, 2-4, 3-3 and 3-2) directed to conformational epitopes of FMDV A/Arg/01 [29]. …”

Section: Methodsmentioning

confidence: 99%

Transient Gene Expression in Serum-Free Suspension-Growing Mammalian Cells for the Production of Foot-and-Mouth Disease Virus Empty Capsids

et al. 2013

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Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. It produces severe economic losses in the livestock industry. Currently available vaccines are based on inactivated FMD virus (FMDV). The use of empty capsids as a subunit vaccine has been reported to be a promising candidate because it avoids the use of virus in the vaccine production and conserves the conformational epitopes of the virus. In this report, we explored transient gene expression (TGE) in serum-free suspension-growing mammalian cells for the production of FMDV recombinant empty capsids as a subunit vaccine. The recombinant proteins produced, assembled into empty capsids and induced protective immune response against viral challenge in mice. Furthermore, they were recognized by anti-FMDV bovine sera. By using this technology, we were able to achieve expression levels that are compatible with the development of a vaccine. Thus, TGE of mammalian cells is an easy to perform, scalable and cost-effective technology for the production of a recombinant subunit vaccine against FMDV.

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Rapid methodology for antigenic profiling of FMDV field strains and for the control of identity, purity and viral integrity in commercial virus vaccines using monoclonal antibodies

Cited by 27 publications

References 32 publications

Early Adaptive Immune Responses in the Respiratory Tract of Foot-and-Mouth Disease Virus-Infected Cattle

Early Adaptive Immune Responses in the Respiratory Tract of Foot-and-Mouth Disease Virus-Infected Cattle

Characterization of monoclonal antibodies against foot-and-mouth disease virus serotype O and application in identification of antigenic variation in relation to vaccine strain selection

Transient Gene Expression in Serum-Free Suspension-Growing Mammalian Cells for the Production of Foot-and-Mouth Disease Virus Empty Capsids

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